アプリケーション

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション

Abreviews

特記事項

Flow Cyt

Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB

Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 80 kDa). For optimal results, primary antibody incubations should be performed at room temperature.

ICC/IF

1/200.

ターゲット情報

機能Phosphorylates inhibitors of NF-kappa-B thus leading to the dissociation of the inhibitor/NF-kappa-B complex and ultimately the degradation of the inhibitor. May play a special role in the immune response. Protects cells against DNA damage-induced cell death.

Anti-IKKi/IKKe antibody [72B587] 画像

ICC/IF image of ab12142 stained Jeg3 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab12142, 1/200 dilution) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM

IKKi/IKKen (80 kDa) detection by Western blot. The analysis of 10 µg of total cell extract from Daudi cells with the anti-IKK iota/IKK epsilon at 0.5 µg/ml (lane 1) and 3 µg/ml (lane 2) dilution.

Flow Cytometry-Anti-IKKi/IKKe antibody [72B587](ab12142)

Overlay histogram showing Jurkat cells stained with ab12142 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab12142, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.